CN116410875B - Rhodotorula megaterium for producing oligosaccharide prebiotics and application thereof - Google Patents
Rhodotorula megaterium for producing oligosaccharide prebiotics and application thereof Download PDFInfo
- Publication number
- CN116410875B CN116410875B CN202310195581.8A CN202310195581A CN116410875B CN 116410875 B CN116410875 B CN 116410875B CN 202310195581 A CN202310195581 A CN 202310195581A CN 116410875 B CN116410875 B CN 116410875B
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- CN
- China
- Prior art keywords
- soy sauce
- rhodotorula
- rhodotorula glutinis
- fermentation production
- production process
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 241000223252 Rhodotorula Species 0.000 title claims description 6
- 235000013406 prebiotics Nutrition 0.000 title abstract description 17
- 229920001542 oligosaccharide Polymers 0.000 title abstract description 15
- 150000002482 oligosaccharides Chemical class 0.000 title abstract description 15
- 235000013555 soy sauce Nutrition 0.000 claims abstract description 46
- 241000223253 Rhodotorula glutinis Species 0.000 claims abstract description 34
- 238000000855 fermentation Methods 0.000 claims abstract description 27
- 230000004151 fermentation Effects 0.000 claims abstract description 26
- 238000004519 manufacturing process Methods 0.000 claims abstract description 14
- 241000007098 Rhodotorula dairenensis Species 0.000 claims abstract description 11
- 239000000796 flavoring agent Chemical class 0.000 claims abstract description 10
- 235000019634 flavors Nutrition 0.000 claims abstract description 10
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- 150000002632 lipids Chemical class 0.000 claims abstract description 5
- 230000002503 metabolic effect Effects 0.000 claims abstract description 5
- 238000001556 precipitation Methods 0.000 claims abstract description 5
- YGHRJJRRZDOVPD-UHFFFAOYSA-N 3-methylbutanal Chemical compound CC(C)CC=O YGHRJJRRZDOVPD-UHFFFAOYSA-N 0.000 claims description 4
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 claims description 4
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 claims description 4
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/165—Yeast isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/50—Soya sauce
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention discloses a rhodotorula glutinis strain with high yield of oligosaccharide prebiotics and application thereof, wherein the rhodotorula glutinis strain (Rhodotoruladairenensis) GT_C14 is preserved in China center for type culture collection (CCTCCNO: M20222011) in the year 2022 and the month 23, the preservation address is China university of Wuhan and Wuhan, the rhodotorula glutinis strain has good stress resistance, and the rhodotorula glutinis strain is added in the soy sauce fermentation production process to high yield of oligosaccharide prebiotics, up-regulate the diversity, the richness and the metabolic activity of lactic acid bacteria and microzyme, enrich short-chain fatty acid and flavor derivatives thereof, inhibit soy sauce precipitation and lipid rancidity, prolong the shelf life of soy sauce, and has good application prospect.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to rhodotorula glutinis capable of producing oligosaccharide prebiotics and application thereof.
Background
Soy sauce is a seasoning prepared from soybean, wheat, etc. by fermentation of microorganism. In addition to the flavoring effect, consumers are increasingly focusing on the probiotic properties of food products. Common oligosaccharides, such as sucrose, maltose, etc., which are commonly found in soy sauce, are often digested and absorbed, and energy metabolized, rather than exhibiting prebiotic properties.
Prebiotic oligosaccharides such as fructo-oligosaccharides, isomaltooligosaccharides and the like are rare in soy sauce, and the prebiotic oligosaccharides are used as natural sweeteners and have the characteristics of mildness, fineness and capability of shielding peculiar smell, which are not possessed by sucrose, white granulated sugar or other sweeteners commonly used in soy sauce preparation. In addition, the prebiotics play a role in regulating and controlling flora in the soy sauce mash fermentation, the core fermentation probiotics and saccharomycetes are subjected to the gain effect, the stress resistance and the metabolic capacity are enhanced, and the comprehensive quality of the soy sauce is improved.
At present, no report on a core brewing strain capable of high yield of prebiotics, regulation of brewing microecology and improvement of taste and flavor of soy sauce exists in the field.
Disclosure of Invention
This section is intended to outline some aspects of embodiments of the application and to briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section as well as in the description of the application and in the title of the application, which may not be used to limit the scope of the application.
The present invention has been made in view of the above and/or problems occurring in the prior art.
Therefore, the invention aims to overcome the defects in the prior art and provide the rhodotorula glutinis (Rhodotorula dairenensis) GT_C14 which is preserved in China center for type culture Collection (CCTCC NO: M20222011) at the 12 th month 23 of 2022 and has the preservation address of university of China, wuhan and Wuhan.
As a preferable scheme of the rhodotorula glutinis, the invention comprises the following steps: the rhodotorula glutinis grows in an environment with 20% of NaCl w/w and pH=4 and 6% of ethanol w/w, and has good stress resistance.
It is a further object of the present invention to overcome the deficiencies of the prior art and to provide the use of rhodotorula glutinis (Rhodotorula dairenensis) GT_C14 in soy fermentation production.
As a preferred embodiment of the application according to the invention, wherein: comprises adding rhodotorula glutinis in the soy sauce fermentation production process to produce oligosaccharide prebiotics with high yield.
As a preferred embodiment of the application according to the invention, wherein: the rhodotorula glutinis serves as a sweet taste regulator in the soy sauce fermentation production process.
As a preferred embodiment of the application according to the invention, wherein: comprises adding rhodotorula glutinis in soy sauce fermentation production process, and up-regulating diversity, richness and metabolic activity of lactobacillus and saccharomycetes.
As a preferred embodiment of the application according to the invention, wherein: comprises adding rhodotorula glutinis into soy sauce fermentation production process, and enriching short chain fatty acid and flavor derivative thereof.
As a preferred embodiment of the application according to the invention, wherein: comprises adding rhodotorula glutinis during soy sauce fermentation production process, inhibiting soy sauce precipitation and lipid rancidity, and prolonging shelf life of soy sauce.
The invention has the beneficial effects that:
The invention provides rhodotorula glutinis (Rhodotorula dairenensis) GT_C14, which is preserved in China Center for Type Culture Collection (CCTCC) No. M20222011 at the 12 month 23 day 2022, and has the preservation address of China university of Wuhan and Wuhan, wherein the strain grows in the environment with 20% of NaCl w/w, pH=4 and 6% of ethanol w/w, has good stress resistance, is added in the soy sauce fermentation production process, has high yield of oligosaccharide prebiotics, up-regulates the diversity, the richness and the metabolic activity of lactobacillus and microzyme, enriches short-chain fatty acid and flavor derivatives thereof, inhibits soy sauce precipitation and lipid rancidity, prolongs the shelf life of soy sauce, and has good application prospect.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art. Wherein:
FIG. 1 is a colony morphology of Rhodotorula megaterium on YPD nutrient agar and an optical microscopy image in an example of the invention.
FIG. 2 is a graph showing stress growth data of rhodotorula glutinis according to an embodiment of the present invention.
FIG. 3 is a graph showing comparison of microbial populations under the intervention of rhodotorula glutinis in the examples of the present invention.
FIG. 4 is a comparative diagram of deterioration of soy sauce in the examples of the present invention.
Detailed Description
In order that the above-recited objects, features and advantages of the present invention will become more apparent, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways other than those described herein, and persons skilled in the art will readily appreciate that the present invention is not limited to the specific embodiments disclosed below.
Further, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic can be included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
The invention provides a rhodotorula glutinis (Rhodotorula dairenensis) GT_C14 which is preserved in China Center for Type Culture Collection (CCTCC) No. M20222011 in 2022 and has a preservation address of China university of Wuhan.
Example 1: separation of rhodotorula glutinis
The soy sauce mash is prepared by ageing high-salt diluted soy sauce mash from Tianjin university for 3 years, sampling by adopting a five-point method, fully and uniformly mixing, placing in a sterile glycerin EP pipe, and storing in a refrigerator at-80 ℃.
Under aseptic condition, weighing 5.0g of soy sauce sample stored at-80deg.C, mixing with 45mL of sterile physiological saline, shaking in a shaker for 2 hr to disperse microbial cells, making into 10 -1 dilution, and performing serial gradient dilution. 0.1mL of dilutions 10 -3、10-4 and 10 -5 are respectively taken and inoculated into YPD nutrient agar medium, the plates are respectively placed in a 30 ℃ incubator for culturing 48 hours after coating, colonies generating pigment are observed and picked, and repeated streaking and purification are carried out to obtain pure strain.
Plate streaking is carried out on YPD solid culture medium, the culture is carried out for 48 hours in a constant temperature incubator at 30 ℃, and the colony morphology is observed: the size, color, shape of colonies, edges of colonies, raised condition, as shown in FIG. 1.
It can be seen that the color of the picked colonies appears orange; the appearance is mainly round, the surface is moist and sticky, the appearance is easy to pick up, the surface is smooth, and the throwing spores are generated.
The screened strain can not ferment galactose, maltose, sucrose, melibiose, lactose, cellobiose and starch; can assimilate glucose, galactose, sorbose, ribose, xylose, L-arabinose, D-arabinose, sucrose, maltose, cellobiose. The optimal growth temperature is 30+/-2 ℃ and the highest temperature is 45 ℃.
Example 2: molecular biological identification of rhodotorula glutinis
Colonies with the size of small rice grains (with the clone diameter of 0.1-0.3 cm) are picked into TE solution, and any solid culture medium (mixed with the culture medium can influence the subsequent genome extraction effect) is avoided as much as possible.
100. Mu.L of TE solution was added, the pellet was resuspended sufficiently, and 20. Mu.L of lysozyme and 10. Mu.L of RNase A were added, mixed thoroughly, incubated at 37℃for 2.5 hours, and shaken once every 30 minutes. 100. Mu.L LYS solution (preheated in advance at 55deg.C) was added and gently mixed by blowing. Then, 20. Mu.L of proteinase K solution was added thereto and the mixture was thoroughly mixed. Incubate at 55℃for 30 min. 220. Mu.L of EXT solution was added. The mixture was inverted several times until it was mixed, and centrifuged at 10,000Xg for 5 minutes. The supernatant was transferred to a fresh 1.5mL EP tube and 220. Mu.L absolute ethanol was added. The mixture was inverted several times until it was mixed well.
The DNA centrifugation column was loaded into a 2mL collection tube, and the whole sample was transferred to the DNA centrifugation column and centrifuged at 10,000Xg for 1 minute. The waste liquid was discarded and the DNA centrifugation column was refilled into a 2mL collection tube. mu.L of PD solution was added, centrifuged at 10,000Xg for 1 minute, the waste solution was discarded, and the DNA centrifugation column was reloaded into a 2mL collection tube. mu.L of PW solution was added, centrifuged at 10,000Xg for 1 min, and the waste solution was discarded. The DNA centrifugation column was reloaded into a 2mL collection tube and this step was repeated. Centrifuge at 12,000rpm for 2 minutes.
The DNA centrifugation column was reloaded into a clean 1.5mL EP tube and allowed to air dry for 5 minutes at room temperature. Add 50-100. Mu.L of the Elution Buffer or water to the DNA spin column. Standing for 3-5 minutes at room temperature. The genomic DNA was collected by centrifugation at 12,000rpm for 2 minutes.
The DNA was stored at-20 ℃. The general primer ITS1: TCCGTAGGTGAACCTGCGG; ITS4: TCCTCCGCTTATTGATATGC.
The extracted DNA sample is diluted in proper amount and used as PCR template for amplification with 1×TSE101 gold plate mix of the family Praeparatae. The sequencing results were spliced with Contigexpress and the two-terminal inaccurate portions were removed. The spliced sequences are aligned in NCBI database (blast. NCBI. Lm. Nih. Gov), and the identification result is rhodotorula glutinis (Rhodotorula dairenensis).
The nucleotide sequence (seq_1) of rhodotorula glutinis in the invention is:
GATATGCTTAAGTTCAGCGGGTAGCCCTACCTGATTTCAGGTCAAAGTCAAAAGGTGCGCGATGGCAGGTTTTGAGCAGTCATCACCACAAGGAGAGACGAAACTTATTACGTCTAACACTGATGTGGGATGTTCCACTAAGTCATTTGAGGTGAGCCATTGCTGGCAGACACCCAAGTCCAAGCCTAACATGTTCAGAAAACATATTAGATTGAGATTTCATGACACTGAAACAGGCATGCCTTTCGGAATACCAAAAGGCGCAAGGTGCGTTCAAAGATTCGATGATTCACTGAATTCTGCAATTCACATTACTTATCGCATTTCGCTGCGTTCTTCATCGATGCGAGAGCCAAGAGATCCGTTGTTAAAAGTTTTGTTTAATTAAGATACTCTTACGTTCGTTACACTGATGTTTGATTATTAACCCGGAGGTCAACGGTTCACAGAGGTGGTAGAATCTGATAAGGTCTTTCGACCAATCAATAATGATCCTTCCGCAGG
Example 3: stress resistance determination of rhodotorula glutinis
2ML of the bacterial liquid frozen at-80 ℃ is poured into a 100mLYPD liquid culture medium (2% soybean peptone, 2% glucose, 1% yeast powder, pH=7 adjustment), the cultured bacterial liquid is inoculated into 100mL of fresh YPD liquid according to the proportion of 2% after culturing for 48 hours by a shaking table at 30 ℃ and culturing to OD 600=0.7 by a shaking table at 120rpm at 30 ℃, and the activation is considered to be completed.
The activated strain was inoculated in batches at an inoculum size of 2% to YPD liquid medium containing 0%, 4%, 8%, 12%, 16%, 20% NaCl (w/w); YPD liquid medium at pH 4, 4.5, 5, 5.5, 6, 6.5, 7; and YPD liquid medium with ethanol content (w/w) of 0%, 2%, 4% and 6%. And culturing at 30 ℃ for 120 hours, centrifugally collecting thalli by the culture solution, drying in a baking oven at 55 ℃ for 24 hours until the weight is constant, and reflecting biomass by dry weight.
The measurement results are shown in FIG. 2, and considering that rhodotorula glutinis needs to play a practical role in soy sauce fermentation, the rhodotorula glutinis stress resistance is evaluated by taking biomass of more than 0.01g/L as a threshold value. Tolerance limit thresholds for NaCl, pH and ethanol were found to be 20% (w/w), 4 and 6% (w/w), respectively.
Example 4: oligosaccharide prebiotic production capability of rhodotorula glutinis
2ML of the bacterial liquid frozen at-80 ℃ is poured into a 100mLYPD liquid culture medium, the bacterial liquid is cultured for 48 hours at 30 ℃ by a shaking table at 120rpm, then the cultured bacterial liquid is inoculated into 100mL of fresh YPD liquid according to the proportion of 2 percent, and the bacterial liquid is cultured at 30 ℃ by the shaking table at 120rpm until the OD 600=0.7, thus the activation is considered to be completed.
The activated strain was inoculated into moromi medium (Daqu: 20% saline=1:2.5 w/w) at an inoculum size of 2% (v/v), and cultured at 30℃for 15 days. The fermentation liquor is extruded and filtered for 3 times by a plate frame, and 10000 Xg of supernatant is taken for freeze-drying and then dissolved by distilled water solution.
The above procedure was repeated with the standard deposited strain Rhodotorula DAIRENENSIS NCYC 3725 (https:// www.mingzhoubio.com /) as a control group to verify the ability to produce oligosaccharide prebiotics.
Determination of ketose content by High Performance Liquid Chromatography (HPLC): a four-stage pump (Delta 600, waters, milford) was coupled to LunaNH 2 columns (4.6X1250 mm) and to pre-column-NH 2 (Phenomnex, torrane). The column temperature was maintained at 30 ℃. The light scattering detector ELSD (mod.1000) was used, balanced at 40 ℃. An automatic syringe (model 717plus, waters, milford, ct, usa) was used with an injection volume of 10 μl. The analyte was eluted with acetonitrile/water and degassed with helium at a flow rate of 1.0mL/min for 35 minutes (acetonitrile: water=77:235 minutes; 73:277 minutes; 73:2714 minutes; 77:23 greater than 1 minute and held until the end of the analysis). The obtained data were analyzed using Waters Millennium software. The fructooligosaccharides content was analyzed by ion chromatography. Ion chromatography conditions: carboPakTM PA200 chromatography column (250 mm. Times.4 mm), carboPakTMPA10 guard column (50 mm. Times.4 mm); column temperature is 30 ℃; the flow rate is 1mL/min, and the sample injection amount is 25 mu L. Gradient elution conditions: 0 to 5min,100mmol/L sodium hydroxide, 0 to 120mmol/L sodium acetate; 5-50 min,100mmol/L sodium hydroxide, 0-120 mmol/L sodium acetate; 10 to 50min,100mmol/L sodium hydroxide, 120 to 320mmol/L sodium acetate; 50-55 min,100mmol/L sodium hydroxide and 320mmol/L sodium acetate; 55-65 min,100mmol/L sodium hydroxide. High performance liquid chromatography to determine trehalose content: mobile phase: acetonitrile: water (v/v) =70:30; flow rate: 1mL/min; column temperature: 25 ℃; chromatographic column: NH 2 bonded phase column, 250mm 4.6mm; a detector: differential refractive light detector. Quantification of the compounds was based on their peak area versus standard.
The fermentation yields oligosaccharide prebiotics are shown in table 1.
TABLE 1 comparison of oligosaccharide prebiotic yield
Compared with the standard strain, the strain of the invention has the advantages that the total ketose, fructo-oligosaccharide and trehalose in the fermentation broth are respectively improved by 6.17 times, 3.67 times and 19.78 times.
Example 5: application of rhodotorula glutinis in soy sauce brewing and application thereof
2ML of the bacterial liquid frozen at-80 ℃ is poured into a 100mLYPD liquid culture medium, the bacterial liquid is cultured for 48 hours at 30 ℃ by a shaking table at 120rpm, then the cultured bacterial liquid is inoculated into 100mL of fresh YPD liquid according to the proportion of 2 percent, and the bacterial liquid is cultured at 30 ℃ by the shaking table at 120rpm until the OD 600=0.7, thus the activation is considered to be completed.
Soaking soybean for 4 hr, sterilizing with parched semen Tritici Aestivi under high pressure steam, continuously steaming for 10min, and maintaining high pressure (0.8 MPa) for 20min. After the temperature was lowered to 37 ℃, 0.3% of starter culture was added to the raw material and incubated at 30 ℃. Turning over at 8h and 12h, and preparing Daqu after 36 h. Mixing Daqu and 18% (w/w) NaCl solution at a ratio of 1:2.5, supplementing activated rhodotorula mucilaginosa with fermentation raw material of 0.5% by mass, and starting saline fermentation. Fermenting at 15deg.C for 30 days, then heating to 30deg.C, fermenting for 30 days, continuously supplementing Rhodotorula mucilaginosa with fermentation raw material of 0.1% by mass, and fermenting at thermal insulation for 120 days. Filtering to obtain soy sauce by plate frame squeezing filtration.
The same mass of zygosaccharomyces rouxii (zygosaccharomyces rouxii Zygosaccharomyces rouxii CICC, purchased from China industry microbiological culture Collection center) was added to the control group, and the same mass of standard deposited strain Rhodotorula DAIRENENSIS NCYC 3725 (https:// www.mingzhoubio.com) was added to the control group, and the other processes were the same as described above.
And (3) detecting soy sauce fermentation flora: extracting genomic DNA of a sample by an SDS method, diluting the sample after quality inspection for PCR amplification, recovering the quality control of a PCR product, constructing a library, and adopting NovaSeq6000 for high-throughput sequencing. Sequencing raw data were quality controlled, clustered with 97% identity, and analyzed for species annotation with the SSUrRNA database of SILVA132 using the Mothur method.
And detecting the volatile flavor substances of the experimental group and the control group by using a headspace solid-phase microextraction and gas chromatography-mass spectrometry combined instrument. 3mL of the post-fermentation sample was placed in a headspace bottle, and 30uL of 20ppm 2-octanol standard was added. After sealing the headspace bottle, the headspace bottle was equilibrated in a heated magnetic stirrer in a 60 ℃ water bath for 20min. After equilibration, the headspace was taken for extraction for 30min at 60℃and desorption for 15min at 250℃with GC-MS injection. GC conditions: the initial temperature was maintained at 40℃for 3min, at 4℃per min up to 150℃for 1min, and at 8℃per min up to 250℃for 6min. Helium is used as carrier gas, the temperature of the sample inlet is 250 ℃, and the sample injection time is 1min. MS conditions: the ion source temperature is 200 ℃, the interface temperature is 220 ℃, the solvent delay time is 1.5min, the ionization mode is EI, the scanning mass range of 70eV is 33-500 m/z, and the scanning speed is 3.00s cans/s. And searching and selecting the compounds with the similarity more than or equal to 90 percent according to the NIST11 database, and combining ion fragments and retention indexes for qualitative determination. 2-octanol is used as an internal standard substance, and an internal standard method is adopted for quantification.
Sealing and packaging the fermented soy sauce, exposing natural environment for 12 months, and taking out the sensory. With reference to GB/T18186-2000, a soy sauce sensory evaluation table was formulated. 10 trained panelists were then selected to evaluate the flavor, color and texture of the soy sauce samples. And team members received several exercises, including definitions and descriptions, prior to formal sensory evaluation. A proper amount of evenly mixed samples are taken and placed in a white porcelain plate with the diameter of 60-90 mm, and sensory is carried out under natural light (based on the instant soy sauce of the same method, scoring is carried out according to the deterioration conditions of fragrance, color and sediment, and no difference is 0-10 minutes of strong difference).
The soy brewing flora (in the moromi fermented for 60 days) is shown in FIG. 3. The conventional soy sauce flora is dominated by bacillus subtilis and Weissella, generates resource competition and has poor fragrance producing capability. After treatment of rhodotorula glutinis gt_c14, based on the probiotic, stress-resistant effects of the prebiotics, the biomass of lactic acid bacteria (tetracoccus halophilus, lactobacillus plantarum, lactobacillus acidophilus) and yeasts (zygosaccharomyces rouxii and wegener's) was significantly improved, the calculated aroma index (representing the diversity of the flora) and chao index (representing the abundance of the flora) were as high as 4.37 and 841, respectively, whereas the standard strain group was only 3.14 and 569. The flora intervention ability of Rhodotorula glutinis GT_C14 is significantly higher than that of the standard deposited strain Rhodotorula DAIRENENSIS NCYC 3725. The soy sauce brewing flavor components are shown in table 2.
TABLE 2 comparison of flavor components of soy brewing
As can be seen, the short chain fatty acids propionic acid and isovaleric acid of the soy sauce of the rhodotorula glutinis GT_C14 fermentation experimental group respectively improve 514.03 mug/L and 393.73 mug/L relative to the control group, and acetic acid reduces 2752.53 mug/L, so that the quality of the soy sauce is prevented from being covered by the over-strong sour taste. In addition, the isoamyl alcohol, the isovaleraldehyde and the 3-methylthiopropanal which are short-chain fatty acid derivatives are respectively improved by 10.89 times, 25.04 times and 31.90 times, and the characteristic aroma of the corresponding malt aroma, caramel aroma, boiled potatoes and the like is obviously improved.
As shown in FIG. 4, the soy sauce deterioration was found to be slight in precipitation and color deterioration without spoilage taste perceived by the soy sauce of the Rhodotorula glutinosa GT-C14 fermentation test group. The three indexes of the first control group are seriously deteriorated, and the quality control capability of the standard preservation strain Rhodotorula DAIRENENSIS NCYC 3725,3725 is obviously lower than that of the strain of the invention.
The strain of the invention has high-yield oligosaccharide, especially trehalose, plays a main role in preventing protein denaturation, inhibiting lipid oxidative deterioration and flavoring; the fructo-oligosaccharide prolongs the shelf life of the product and increases the viscosity and the heavy feel of the product; the ketose has mild and fine sweet taste, can mask peculiar smell (beany flavor and oily fishy smell), and can maintain antioxidant level of soy sauce for a longer time.
It should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted without departing from the spirit and scope of the technical solution of the present invention, and it should be covered in the scope of the present invention.
Claims (6)
1. Rhodotorula megaterium (Rhodotorula dairenensis) GT_C14 was deposited at China center for type culture Collection (CCTCC NO: M20222011) at 12/23 of 2022 with a deposit address of university of Chinese, wuhan, and Wuhan.
2. Use of rhodotorula glutinis according to claim 1 in soy sauce fermentation production.
3. The use according to claim 2, wherein: the rhodotorula glutinis serves as a sweet taste regulator in the soy sauce fermentation production process.
4. The use according to claim 2, wherein: comprises adding rhodotorula glutinis in soy sauce fermentation production process, and up-regulating diversity and metabolic activity of lactobacillus and saccharomycetes.
5. The use according to claim 2, wherein: comprises adding Rhodotorula megaterium in soy sauce fermentation production process, and enriching short-chain fatty acid and its flavor derivatives, wherein the short-chain fatty acid is acid propionic acid and isovaleric acid, and the short-chain fatty acid derivatives are isoamyl alcohol, isovaleraldehyde, and 3-methylthiopropanal.
6. The use according to claim 2, wherein: comprises adding rhodotorula glutinis during soy sauce fermentation production process, inhibiting soy sauce precipitation and lipid rancidity, and prolonging shelf life of soy sauce.
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